ILVC_LACLA
ID ILVC_LACLA Reviewed; 340 AA.
AC Q02138; Q9CG82;
DT 01-JUL-1993, integrated into UniProtKB/Swiss-Prot.
DT 29-AUG-2001, sequence version 2.
DT 03-AUG-2022, entry version 141.
DE RecName: Full=Ketol-acid reductoisomerase (NADP(+)) {ECO:0000255|HAMAP-Rule:MF_00435, ECO:0000303|PubMed:23776225};
DE Short=KARI {ECO:0000255|HAMAP-Rule:MF_00435, ECO:0000303|PubMed:23776225};
DE EC=1.1.1.86 {ECO:0000255|HAMAP-Rule:MF_00435, ECO:0000269|PubMed:23776225};
DE AltName: Full=Acetohydroxy-acid isomeroreductase {ECO:0000255|HAMAP-Rule:MF_00435};
DE Short=AHIR {ECO:0000255|HAMAP-Rule:MF_00435};
DE AltName: Full=Alpha-keto-beta-hydroxylacyl reductoisomerase {ECO:0000255|HAMAP-Rule:MF_00435};
DE AltName: Full=Ketol-acid reductoisomerase type 1 {ECO:0000255|HAMAP-Rule:MF_00435, ECO:0000303|PubMed:23776225};
DE AltName: Full=Ketol-acid reductoisomerase type I {ECO:0000255|HAMAP-Rule:MF_00435, ECO:0000303|PubMed:23776225};
GN Name=ilvC {ECO:0000255|HAMAP-Rule:MF_00435}; OrderedLocusNames=LL1226;
GN ORFNames=L0080;
OS Lactococcus lactis subsp. lactis (strain IL1403) (Streptococcus lactis).
OC Bacteria; Firmicutes; Bacilli; Lactobacillales; Streptococcaceae;
OC Lactococcus.
OX NCBI_TaxID=272623;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND PATHWAY.
RC STRAIN=NCDO 2118;
RX PubMed=1400210; DOI=10.1128/jb.174.20.6580-6589.1992;
RA Godon J.-J., Chopin M.-C., Ehrlich S.D.;
RT "Branched-chain amino acid biosynthesis genes in Lactococcus lactis subsp.
RT lactis.";
RL J. Bacteriol. 174:6580-6589(1992).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=IL1403;
RX PubMed=11337471; DOI=10.1101/gr.gr-1697r;
RA Bolotin A., Wincker P., Mauger S., Jaillon O., Malarme K., Weissenbach J.,
RA Ehrlich S.D., Sorokin A.;
RT "The complete genome sequence of the lactic acid bacterium Lactococcus
RT lactis ssp. lactis IL1403.";
RL Genome Res. 11:731-753(2001).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP MUTAGENESIS OF VAL-48; ARG-49; LYS-52 AND SER-53.
RX PubMed=23776225; DOI=10.1073/pnas.1306073110;
RA Brinkmann-Chen S., Flock T., Cahn J.K., Snow C.D., Brustad E.M.,
RA McIntosh J.A., Meinhold P., Zhang L., Arnold F.H.;
RT "General approach to reversing ketol-acid reductoisomerase cofactor
RT dependence from NADPH to NADH.";
RL Proc. Natl. Acad. Sci. U.S.A. 110:10946-10951(2013).
CC -!- FUNCTION: Involved in the biosynthesis of branched-chain amino acids
CC (BCAA). Catalyzes an alkyl-migration followed by a ketol-acid reduction
CC of (S)-2-acetolactate (S2AL) to yield (R)-2,3-dihydroxy-isovalerate. In
CC the isomerase reaction, S2AL is rearranged via a Mg-dependent methyl
CC migration to produce 3-hydroxy-3-methyl-2-ketobutyrate (HMKB). In the
CC reductase reaction, this 2-ketoacid undergoes a metal-dependent
CC reduction by NADPH to yield (R)-2,3-dihydroxy-isovalerate.
CC {ECO:0000255|HAMAP-Rule:MF_00435, ECO:0000269|PubMed:23776225}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(2R)-2,3-dihydroxy-3-methylbutanoate + NADP(+) = (2S)-2-
CC acetolactate + H(+) + NADPH; Xref=Rhea:RHEA:22068, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:49072, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349,
CC ChEBI:CHEBI:58476; EC=1.1.1.86; Evidence={ECO:0000255|HAMAP-
CC Rule:MF_00435, ECO:0000269|PubMed:23776225};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(2R,3R)-2,3-dihydroxy-3-methylpentanoate + NADP(+) = (S)-2-
CC ethyl-2-hydroxy-3-oxobutanoate + H(+) + NADPH; Xref=Rhea:RHEA:13493,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:49256, ChEBI:CHEBI:49258,
CC ChEBI:CHEBI:57783, ChEBI:CHEBI:58349; EC=1.1.1.86;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_00435};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_00435};
CC Note=Binds 2 magnesium ions per subunit. {ECO:0000255|HAMAP-
CC Rule:MF_00435};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=13 uM for NADPH (at pH 7) {ECO:0000269|PubMed:23776225};
CC KM=285 uM for NADH (at pH 7) {ECO:0000269|PubMed:23776225};
CC Note=kcat is 0.8 sec(-1) for reductoisomerase activity with NADPH as
CC substrate (at pH 7). kcat is 0.1 sec(-1) for reductoisomerase
CC activity with NADH as substrate (at pH 7).
CC {ECO:0000269|PubMed:23776225};
CC -!- PATHWAY: Amino-acid biosynthesis; L-isoleucine biosynthesis; L-
CC isoleucine from 2-oxobutanoate: step 2/4. {ECO:0000255|HAMAP-
CC Rule:MF_00435, ECO:0000305|PubMed:1400210}.
CC -!- PATHWAY: Amino-acid biosynthesis; L-valine biosynthesis; L-valine from
CC pyruvate: step 2/4. {ECO:0000255|HAMAP-Rule:MF_00435,
CC ECO:0000305|PubMed:1400210}.
CC -!- SIMILARITY: Belongs to the ketol-acid reductoisomerase family.
CC {ECO:0000255|HAMAP-Rule:MF_00435}.
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DR EMBL; U92974; AAB81921.1; -; Genomic_DNA.
DR EMBL; AE005176; AAK05324.1; -; Genomic_DNA.
DR PIR; B86778; B86778.
DR PIR; S35140; S35140.
DR RefSeq; NP_267382.1; NC_002662.1.
DR RefSeq; WP_003131136.1; NC_002662.1.
DR AlphaFoldDB; Q02138; -.
DR SMR; Q02138; -.
DR STRING; 272623.L0080; -.
DR PaxDb; Q02138; -.
DR EnsemblBacteria; AAK05324; AAK05324; L0080.
DR KEGG; lla:L0080; -.
DR PATRIC; fig|272623.7.peg.1325; -.
DR eggNOG; COG0059; Bacteria.
DR HOGENOM; CLU_033821_0_1_9; -.
DR OMA; RAMFSWL; -.
DR BRENDA; 1.1.1.383; 2864.
DR UniPathway; UPA00047; UER00056.
DR UniPathway; UPA00049; UER00060.
DR Proteomes; UP000002196; Chromosome.
DR GO; GO:0004455; F:ketol-acid reductoisomerase activity; IEA:UniProtKB-UniRule.
DR GO; GO:0000287; F:magnesium ion binding; IEA:UniProtKB-UniRule.
DR GO; GO:0050661; F:NADP binding; IEA:InterPro.
DR GO; GO:0009097; P:isoleucine biosynthetic process; IEA:UniProtKB-UniRule.
DR GO; GO:0009099; P:valine biosynthetic process; IEA:UniProtKB-UniRule.
DR HAMAP; MF_00435; IlvC; 1.
DR InterPro; IPR008927; 6-PGluconate_DH-like_C_sf.
DR InterPro; IPR013023; KARI.
DR InterPro; IPR000506; KARI_C.
DR InterPro; IPR013116; KARI_N.
DR InterPro; IPR014359; KARI_prok.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR PANTHER; PTHR21371; PTHR21371; 1.
DR Pfam; PF01450; IlvC; 1.
DR Pfam; PF07991; IlvN; 1.
DR PIRSF; PIRSF000116; IlvC_gammaproteo; 1.
DR SUPFAM; SSF48179; SSF48179; 1.
DR SUPFAM; SSF51735; SSF51735; 1.
DR TIGRFAMs; TIGR00465; ilvC; 1.
DR PROSITE; PS51851; KARI_C; 1.
DR PROSITE; PS51850; KARI_N; 1.
PE 1: Evidence at protein level;
KW Amino-acid biosynthesis; Branched-chain amino acid biosynthesis; Magnesium;
KW Metal-binding; NADP; Oxidoreductase; Reference proteome.
FT CHAIN 1..340
FT /note="Ketol-acid reductoisomerase (NADP(+))"
FT /id="PRO_0000151318"
FT DOMAIN 3..182
FT /note="KARI N-terminal Rossmann"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01197"
FT DOMAIN 183..328
FT /note="KARI C-terminal knotted"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01198"
FT ACT_SITE 108
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435"
FT BINDING 26..29
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435"
FT BINDING 49
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435"
FT BINDING 53
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435"
FT BINDING 83..86
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435"
FT BINDING 134
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435"
FT BINDING 191
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435"
FT BINDING 191
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435"
FT BINDING 195
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435"
FT BINDING 227
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435"
FT BINDING 231
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435"
FT BINDING 252
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00435"
FT MUTAGEN 48
FT /note="V->L: Inversion of cofactor specificity from NADPH
FT to NADH. Strong decrease of the affinity for NADPH and 2.5-
FT fold increase of the affinity for NADH. 8-fold decrease of
FT the catalytic efficiency for NADPH and 4-fold increase of
FT the catalytic efficiency for NADH; when associated with P-
FT 49, L-52 and D-53."
FT /evidence="ECO:0000269|PubMed:23776225"
FT MUTAGEN 49
FT /note="R->P: Inversion of cofactor specificity from NADPH
FT to NADH. Strong decrease of the affinity for NADPH and 2.5-
FT fold increase of the affinity for NADH. 8-fold decrease of
FT the catalytic efficiency for NADPH and 4-fold increase of
FT the catalytic efficiency for NADH; when associated with L-
FT 48, L-52 and D-53."
FT /evidence="ECO:0000269|PubMed:23776225"
FT MUTAGEN 52
FT /note="K->L: Inversion of cofactor specificity from NADPH
FT to NADH. Strong decrease of the affinity for NADPH and 2.5-
FT fold increase of the affinity for NADH. 8-fold decrease of
FT the catalytic efficiency for NADPH and 4-fold increase of
FT the catalytic efficiency for NADH; when associated with L-
FT 48, P-49 and D-53."
FT /evidence="ECO:0000269|PubMed:23776225"
FT MUTAGEN 53
FT /note="S->D: Inversion of cofactor specificity from NADPH
FT to NADH. Strong decrease of the affinity for NADPH and 2.5-
FT fold increase of the affinity for NADH. 8-fold decrease of
FT the catalytic efficiency for NADPH and 4-fold increase of
FT the catalytic efficiency for NADH; when associated with L-
FT 48, P-49 and L-52."
FT /evidence="ECO:0000269|PubMed:23776225"
FT CONFLICT 270
FT /note="A -> E (in Ref. 1; AAB81921)"
FT /evidence="ECO:0000305"
FT CONFLICT 303
FT /note="T -> I (in Ref. 1; AAB81921)"
FT /evidence="ECO:0000305"
FT CONFLICT 321..340
FT /note="LRKAMPFTQSGDDDAFKIYQ -> HVKQCHSHNLVMTMPLKSISNFSY (in
FT Ref. 1; AAB81921)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 340 AA; 37211 MW; 93E2FF206D7AFF2F CRC64;
MAVTMYYEDD VEVSALAGKQ IAVIGYGSQG HAHAQNLRDS GHNVIIGVRH GKSFDKAKED
GFETFEVGEA VAKADVIMVL APDELQQSIY EEDIKPNLKA GSALGFAHGF NIHFGYIKVP
EDVDVFMVAP KAPGHLVRRT YTEGFGTPAL FVSHQNASGH AREIAMDWAK GIGCARVGII
ETTFKEETEE DLFGEQAVLC GGLTALVEAG FETLTEAGYA GELAYFEVLH EMKLIVDLMY
EGGFTKMRQS ISNTAEFGDY VTGPRIITDA VKKNMKLVLA DIQSGKFAQD FVDDFKAGRP
KLTAYREAAK NLEIEKIGAE LRKAMPFTQS GDDDAFKIYQ
